Overview

  • Product name

    Anti-Rhodopsin antibody [EPR21876]
    See all Rhodopsin primary antibodies
  • Description

    Rabbit monoclonal [EPR21876] to Rhodopsin
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, IHC-Fr, IPmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Mouse Rhodopsin aa 1 to the C-terminus. The exact sequence is proprietary.
    Database link: P15409

  • Positive control

    • WB: Human, mouse and rat eye tissue lysates; Rat retina lysate. IHC-P: Human, mouse and rat retina tissues. IHC-Fr: Mouse and rat retina. IP: Rat eye tissue lysate.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR21876
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab221664 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/1000. Detects a band of approximately 35-150 kDa (predicted molecular weight: 38 kDa).
IHC-P 1/32000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

(mouse and rat). 1/4000 (human).

IHC-Fr 1/500.

Antigen retrieval is not needed.

IP 1/30.

Target

  • Function

    Photoreceptor required for image-forming vision at low light intensity. Required for photoreceptor cell viability after birth. Light-induced isomerization of 11-cis to all-trans retinal triggers a conformational change leading to G-protein activation and release of all-trans retinal.
  • Tissue specificity

    Rod shaped photoreceptor cells which mediates vision in dim light.
  • Involvement in disease

    Retinitis pigmentosa 4
    Night blindness, congenital stationary, autosomal dominant 1
  • Sequence similarities

    Belongs to the G-protein coupled receptor 1 family. Opsin subfamily.
  • Post-translational
    modifications

    Phosphorylated on some or all of the serine and threonine residues present in the C-terminal region.
    Contains one covalently linked retinal chromophore.
  • Cellular localization

    Membrane. Synthesized in the inner segment (IS) of rod photoreceptor cells before vectorial transport to the rod outer segment (OS) photosensory cilia.
  • Information by UniProt
  • Database links

  • Alternative names

    • CSNBAD1 antibody
    • MGC138309 antibody
    • MGC138311 antibody
    • OPN 2 antibody
    • OPN2 antibody
    • opsd antibody
    • OPSD_HUMAN antibody
    • opsin 2 antibody
    • Opsin 2 rod pigment antibody
    • Opsin-2 antibody
    • Opsin2 antibody
    • Retinitis Pigmentosa 4 antibody
    • Retinitis pigmentosa 4 autosomal dominant antibody
    • RHO antibody
    • Rhodopsin antibody
    • RP 4 antibody
    • RP4 antibody
    see all

Images

  • Immunohistochemical analysis of paraffin-embedded human retina tissue labeling Rhodopsin with ab221664 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining in outer segment of human retina is observed (PMID: 23223288). Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen mouse retina tissue labeling Rhodopsin with ab221664 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Positive staining in the outer segment of mouse retina is observed (PMID: 21938483).

    The nuclear counter stain is DAPI (blue).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

  • Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen rat retina tissue labeling Rhodopsin with ab221664 at 1/500 dilution,followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Positive staining in the outer segment of rat retina is observed (PMID: 21938483).

    The nuclear counter stain is DAPI (blue).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

  • Immunohistochemical analysis of paraffin-embedded mouse retina tissue labeling Rhodopsin with ab221664 at 1/32000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining in outer segment of mouse retina is observed (PMID: 23223288). Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded rat retina tissue labeling Rhodopsin with ab221664 at 1/32000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining in outer segment of rat retina is observed (PMID: 23223288). Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Lanes 1-3 : Anti-Rhodopsin antibody [EPR21876] (ab221664) at 1/1000 dilution
    Lane 4 : Anti-Rhodopsin antibody [EPR21876] (ab221664) at 1/5000 dilution

    Lane 1 : Human eye tissue at 20 µg
    Lane 2 : Mouse eye tissue at 20 µg
    Lane 3 : Rat retina tissue lysate at 20 µg
    Lane 4 : Rat eye tissue lysate at 10 µg

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 38 kDa
    Observed band size: 35-150 kDa
    why is the actual band size different from the predicted?



     

    Exposure Time: Lane 1: 3 minutes; Lane 2: 3 seconds; Lane 3: 58 seconds; Lane 4: 26 seconds.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    The multiple bands (>60 kDa) correspond to dimers and multimers of rhodopsin, consistent with the literature (PMID: 25270370; PMID: 22219383).

     

  • Rhodopsin was immunoprecipitated from 0.35 mg rat eye tissue lysate with ab221664 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab221664 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1/5000 dilution.

    Lane 1: Rat eye tissue lysate 10 μg (input).

    Lane 2: Rat eye tissue lysate (boiled) 10 μg (input).

    Lane 3: ab221664 IP in Rat eye tissue lysate.

    Lane 4: Rabbit monoclonal IgG (ab172730) instead of ab221664 in rat eye tissue lysate.

    Exposure time: 15 seconds

    The multiple bands (>60 kDa) correspond to dimers and multimers of rhodopsin, consistent with the literature (PMID: 25270370; PMID: 22219383).

    We do not recommend boiling samples in loading buffer as this may cause protein aggregation (lane 2).

References

ab221664 has not yet been referenced specifically in any publications.

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